Metabolome Of Rats With Pre-diabetic And Lnsulin Resistance In Intervention Of Gegen Qinlian Decoction

Objective Metabolomics analysis was used to explore the metabolic mechanism of insulin resistance induced by high-fat diet in rats and the effect of Gegen Qinlian Decoction on serum metabolome of pre-diabetic and insulin resistant rats.Trying to clarify the relationship between high-fat diets and obese insulin resistance.And to study the possible mechanism of Gegen Qinlian Decoction for intervening insulin resistance and preventing type 2 diabetes.Methods Insulin resistance(IR)model induced by SD rats with 60% high-fat diet.After successful modeling,all rat serum samples were collected and stored at-80 ° C.The IR group rats were randomly divided into model group and Pueraria The high-,medium-,and low-dose groups of Qinlian Decoction continued to be fed with high-fat diets.In addition,the model group was given intragastric gavage daily.Gegen Qinlian decoction(GQD)was orally administered with crude drugs 14.85,4.85,and 1.65 g / kg.10 ml / kg gavage.GQD was administered for a total of sixteen weeks,and rat serum samples were collected every four weeks,and a total of four times were collected during the administration period.The experimental endpoint was analyzed by UHPLC / Q-TOF-MS Using technology,collect the metabolic profile information of rat serum samples during the modeling end point and during drug administration,and use Mass Profiler Professional(MPP)and Graphpad prism7.0 software to combine METLIN biological database and human metabolite database HMDB for endogenous differential metabolism Analysis and identification.According to the results of the endpoint efficacy index,the metabolome was analyzed in a time-series manner during the administration of the low-and medium-dose administration groups to explore the relevant mechanisms of high-fat diet-induced insulin resistance and the mechanism of GQD intervention in insulin resistance and the prevention of diabetes.Results 1.Twelve weeks after high-fat diet,compared with the normal group,the IR index of the IR group was significantly increased(P <0.05),and there was no difference in blood glucose.The changes of plasma metabolome of the IR group and normal group rats in positive and negative ion mode are as follows,including L-Kynurenine,O-Arachidonoyl,α-Hydroxybutyric acid,and Succinic acid were significantly up-regulated(P <0.05),indole-3-propionic acid,Eicosapentaenoic acid,6,9,12,15,18,21-Tetracosahexaenoic acid were significantly down-regulated(P <0.05).Potential biomarkers are mainly involved in propionate metabolism,butyrate metabolism,tricarboxylic acid cycle,unsaturated fatty acid synthesis,tryptophan cycle and alanine,aspartic acid and glutamic acid metabolism.2.Sixteen weeks after the administration,compared with the normal group,the IR index of the IR group was significantly increased(P <0.01),and the blood glucose was significantly increased(P <0.05).Compared with the IR group,the IR index of the GQD-L group and the GQD-M group was significantly reduced(P <0.05),there was no difference in the GQD-H group,the blood glucose of the GQD-L group was significantly reduced(P <0.01),there was no difference in blood glucose with GQD-M and GQD-H group.Analysis results of serum metabolomics during GQD-L group administration: Deoxycytidine,2-Methyl-3-ketovaleric acid,Xanthosine,and 6,9,12,15,18,21-Tetracosahexaenoic acid were identified at four weeks after administration;γ-Glutamylleucine,2-Methyl-3-ketovaleric acid,LPC,LPE,etc were identified at eight weeks after administration;Leucylproline,LPC were identified at twelve weeks of administration;Hippuric acid,12-Ketodeoxycholic acid,3-Oxo-4,6-choladienoic acid and LPC were identified at the end point.Gegen Qinlian Decoction has different degrees of callback effect on it,which mainly involves metabolic pathways such as amino acid metabolism,phospholipid metabolism,and bile acid metabolism.3.Results of serum metabolome analysis of GQD-M group during administration: Three biomarkers were obtained four weeks after administration,which were 2-Methyl-3-ketovaleric acid,Deoxycytidine,and Xanthosine;seven biomarkers were obtained eight weeks after administration,which were 6,9,12,15,18,21-Tetracosahexaenoic acid,LPE,Sphinganine,etc;twelve weeks after administration,four were obtained,which were 2-Hydroxybutyric acid,2-Methyl-3-ketovaleric acid,Deoxycytidine,and LPC;Five were obtained in the end point,which were Palmitoleic acid,10 E,12Z-Octadecadienoic acid,Docosahexaenoic acid,alpha-Linolenic acid,and Indole-3-propionic acid.GQD-M makes the fatty acid substances have a significant callback effect,which mainly involves the metabolic pathways such as unsaturated fatty acid synthesis and phospholipid metabolism.Conclusion 1.Through data analysis and identification,compared with the normal group,the serum metabolites in the IR group were identified as a total of 7 metabolic differences,including amino acid metabolism,organic acid metabolism,and unsaturated fatty acid biosynthesis.These metabolic pathways are related to inflammation and oxidation.Stress and intestinal flora are closely related,suggesting that long-term high-fat diet or by affecting the intestinal flora and inducing chronic inflammation will further lead to hypoglycemia and decreased insulin sensitivity and eventually develop into insulin resistance and type 2 diabetes.2.Analysis and identification studies have shown that GQD-L administration has a better effect on intervention of insulin resistance and prevention of diabetes,and its mechanism of action may be through intervention of inflammatory response,regulation of intestinal flora,and improvement of bile acid metabolism to play a therapeutic and preventive role.3.GQD-M can regulate insulin resistance in rats,and its mechanism of action is related to fatty acid biosynthesis,phospholipid metabolism,regulation of inflammatory response and oxidative stress response in the body.4.By comparing the results of the serum metabolomics of the two administered doses,the differences between the two groups are mainly concentrated on amino acids and related substances to the intestinal flora,suggesting that the key point for the two doses to exert different effects is on Regulatory effects and effects on intestinal flora.